Dasatinib synergizes with ATRA to trigger granulocytic differentiation in ATRA resistant acute promyelocytic leukemia cell lines via Lyn inhibition-mediated activation of RAF-1/MEK/ERK

Ding, Ming; Weng, Xiang‐Qin; Yan, Sheng; Wu, Jing; Cui, Liang; Cai, Xueli

doi:10.1016/j.fct.2017.10.053

Cited by 9 publications

(8 citation statements)

References 57 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…C/EBPβ, C/EBPε and PU.1 are required for the maturation of the myeloid lineages, as well as ATRA-induced differentiation in APL cells [21][22][23][24]. Moreover, by MEK/ERK modulating the protein levels of C/EBPβ, C/EBPε and PU.1, some medicines including enzastaurin synergize with ATRA to induce differentiation in ATRA-resistant APL cells [16,[25][26][27]. As Fig.…”

Section: Resultsmentioning

confidence: 99%

Enzasataurin Enhances ATRA-induced Differentiation of Acute Myeloid Leukemia Cells

Cui

Ding

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

Abstract Background All-trans retinoic acid (ATRA) is considered to be the sole clinically useful differentiating agent in the treatment of acute myeloid leukemia (AML). However, it has been effective only in acute promyelocytic leukemia (APL) but not other subtypes of AML. Therefore, finding strategies to sensitize cells to ATRA may develop ATRA-based therapy in the treatment of non-APL AML patients. Methods Cell proliferation was assessed by cell growth. Cell death was evaluated by cell viability and Annexin-V assay. Cell differentiation was analyzed by CD11b expression and morphology. To explore the underlying mechanisms, we studied the role of PKCβ, MEK, ERK, AKT, PU.1, C/EBPβ and C/EBPε by Western-blotting analysis. Results In this study, a clinically achievable concentration of enzastaurin enhanced ATRA-induced differentiation of AML cell lines, HL-60 and U937 as well as non-APL AML primary cells, while it also restored ATRA sensitivity in ATRA-resistant cell line, HL-60Res. Mechanistically, in all these cell lines, enzastaurin-ATRA (enz-ATRA) enhanced the protein levels of PU.1, CCAAT/enhancer binding protein β (C/EBPβ) and C/EBPε. The activity of protein kinase C β (PKCβ) was suppressed by enz-ATRA treatment in HL-60 and HL-60Res cells. However, another PKCβ-selective inhibitor mimicked the cellular and molecular effects of enzastaurin only in HL-60 cells. Only in U937 cells, enz-ATRA activated MEK and ERK, and a MEK specific inhibitor suppressed enz-ATRA-triggered differentiation and reduced the protein levels of PU.1, C/EBPβ and C/EBPε. Enz-ATRA activated Akt in HL-60 and HL-60Res cells. However, an Akt inhibitor blocked enz-ATRA-triggered differentiation and restored the protein levels of PU.1, C/EBPβ and C/EBPε only in HL-60Res cells. Therefore, PKCβ inhibition, MEK/ERK and Akt activation are involved in enz-ATRA-induced differentiation in HL-60, U937 and HL-60Res cells, respectively by modulation of the protein levels of C/EBPβ, C/EBPε and PU.1. Conclusions Enzastaurin, at the clinically achievable concentration, enhances ATRA-induced differentiation of AML cells by PKCβ inhibition, MEK/ERK and Akt activation. This study may provide a potential therapeutic strategy for AML patients.

show abstract

Section: Resultsmentioning

confidence: 99%

Enzasataurin Enhances ATRA-induced Differentiation of Acute Myeloid Leukemia Cells

Cui

Ding

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…Many studies suggested that RAF can affect the proliferation of cell through in uencing the distribution of cell cycle [29][30][31], and we speculated that the signi cant and rapid change of estradiol level was caused by GCs proliferation. Our ow cytometry analysis results showed that RAF709 did not induce the suppression of cell cycle stage, but affected on the activity of estradiol synthase, which con rmed that RAF1 could regulate the expression of CYP19A1 [32] through the ERK signaling pathways [33].…”

Section: Discussionmentioning

confidence: 91%

RAF1 as Downstream Molecule Mediates the FSH Signaling Pathway to Stimulate E2 Synthesis and Secretion in Mouse Ovarian Granulosa Cells

Luo

Liu

Guo

et al. 2020

Preprint

View full text Add to dashboard Cite

Background: V-raf-leukemia viral oncogene 1 (RAF1) kinase is the key factor in extracellular signal regulated pathway, which transmits signals to the downstream extracellular regulated protein kinases (ERK). Regulatory function of RAF1 has been proved to mediate steroid hormone synthesis, which played an essential physiological function in reproduction and development. Whether RAF1 takes part in the signaling events of gonadotropic hormones follicle-stimulating hormone (FSH) in ovarian is unknown.Results: We found that RAF1 as downstream molecule mediates the FSH signaling pathway to stimulate estradiol (E2) synthesis and secretion in mouse ovarian granulosa cells (GCs). The expression of RAF1 is induced by FSH and the production of E2 is increased in the serum and primary ovarian GCs supernatant, the process of which is blocked by treating with RAF1 inhibitor (N-(2-Methyl-5'-morpholino-6'-((tetrahydro-2H-pyran-4-yl)oxy)-[3,3'-bipyridin]-5-yl)-3(trifluoromethyl) benzamide, RAF709). Inhibition of RAF1 activity by RAF709 decreased ERK phosphorylation, and suppressed the expression of cytochrome P450 family 19 subfamily a member 1 (CYP19A1) which is a major rate-limiting enzyme to participate in the last step of E2 biosynthesis. Conclusion: Our results suggest that RAF1 play a pivotal mediating roles toward E2 production in FSH signaling pathway by inducing the phosphorylation of ERK and promoting the process of estradiol synthesis. RAF1 may be a potential and effective factor to regulate the function of the female mouse reproductive system.

show abstract

“…Protein lysate preparation and immunoblotting were carried out as previously described [ 13 ]. Briefly, after lysis with RIPA buffer (Beyotime Biotechnology Ltd., Shanghai, China) and centrifugation at 13,000 rpm at 4 ºC for 10 min, the supernatants were harvested, and proteins were quantified by a protein quantification kit (Beyotime Biotechnology Ltd.).…”

Section: Methodsmentioning

confidence: 99%

“…Immunoprecipitation was carried out as previously described [ 13 ]. Briefly, after lysis in RIPA buffer and centrifugation at 13,000 rpm at 4 °C for 10 min, the lysates were incubated separately with 2 μg of the anti-lyn, anti-Fgr, anti-Hck or anti-RAF antibody overnight at 4 °C.…”

Section: Methodsmentioning

confidence: 99%

Combination of midostaurin and ATRA exerts dose-dependent dual effects on acute myeloid leukemia cells with wild type FLT3

et al. 2022

Self Cite

View full text Add to dashboard Cite

Background Midostaurin combined with chemotherapy is currently used to treat newly diagnosed acute myeloid leukemia (AML) patients with FMS-like tyrosine kinase 3 (FLT3)-mutations. However, midostaurin acts as an antagonist to some chemotherapeutic agents in leukemia cell lines without FLT3 mutations. All-trans retinoic acid (ATRA) induces apoptosis when used in combination with midostaurin in FLT3-mutated AML cells. This combination has been shown to be safe in AML patients. However, the effect of this combination has not been investigated in AML without FLT3 mutations. Methods Cell proliferation was assessed by a cell counting assay. Cell death was evaluated by cell viability and Annexin-V assays. Cell differentiation was assessed by CD11b expression profiling and morphological analysis. To explore the underlying mechanisms, we studied the role of caspase3/7, Lyn, Fgr, Hck, RAF, MEK, ERK, AKT, PU.1, CCAAT/enhancer binding protein β (C/EBPβ) and C/EBPε by Western blot analysis and immunoprecipitation assays. Antitumor activity was also confirmed in mouse xenograft models established with AML cells. Results In this study, 0.1 − 0.25 μM midostaurin (mido(L)) combined with ATRA induced differentiation while 0.25 − 0.5 μM midostaurin (mido(H)) combined with ATRA triggered apoptosis in some AML cell lines without FLT3-mutations. Midostaurin combined with ATRA (mido-ATRA) also exhibited antitumor activity in mouse xenograft models established with AML cells. Mechanistically, mido(H)-ATRA-induced apoptosis was dependent on caspase-3/7. Mido(L)-ATRA inhibited Akt activation which was associated with decreased activity of Lyn/Fgr/Hck, resulted in dephosphorylation of RAF S259, activated RAF/MEK/ERK, along with upregulating the protein levels of C/EBPβ, C/EBPε and PU.1. A MEK specific inhibitor was observed to suppress mido(L)-ATRA-induced increases in the protein levels of C/EBPs and PU.1 and mido(L)-ATRA-induced differentiation. Furthermore, inhibition of Akt activity promoted mido(L)-ATRA-induced downregulation of RAF S259 phosphorylation and mido(L)-ATRA-induced differentiation. Therefore, Lyn/Fgr/Hck-associated Akt inhibition activated RAF/MEK/ERK and controlled mido(L)-ATRA-induced differentiation by upregulation of C/EBPs and PU.1. Mido(L)-ATRA also promoted assembly of the signalosome, which may facilitate RAF activation. Conclusions Midostaurin combined with ATRA exerts antitumor activity against AML with wild-type FLT3 mutations in vitro and in vivo. These findings may provide novel therapeutic strategies for some AML patients without FLT3 mutations and imply a new target of midostaurin.

show abstract

Dasatinib synergizes with ATRA to trigger granulocytic differentiation in ATRA resistant acute promyelocytic leukemia cell lines via Lyn inhibition-mediated activation of RAF-1/MEK/ERK

Cited by 9 publications

References 57 publications

Enzasataurin Enhances ATRA-induced Differentiation of Acute Myeloid Leukemia Cells

Enzasataurin Enhances ATRA-induced Differentiation of Acute Myeloid Leukemia Cells

RAF1 as Downstream Molecule Mediates the FSH Signaling Pathway to Stimulate E2 Synthesis and Secretion in Mouse Ovarian Granulosa Cells

Combination of midostaurin and ATRA exerts dose-dependent dual effects on acute myeloid leukemia cells with wild type FLT3

Contact Info

Product

Resources

About